Novel approach to quantitative reverse transcription PCR assay of mRNA component in autopsy material using the TaqMan fluorogenic detection system: dynamics of pulmonary surfactant apoprotein A

A novel approach to quantitative reverse transcription (RT)-PCR assay of mR NA component using fluorescent TaqMan methodology and a new instrument (ABI Prism 7700 sequence detection system) was developed for autopsy materials. Pulmonary surfactant. apoprotein A (SP-A) mRNA from a cadaveric lung was q uantitated in real-time. The target SP-A gene and the endogenous reference of glyceraldehyde-3-phosphate (GAPDH) were amplified in the same tube, and an amount of the target was normalized to the reference. This assay had a h igh reproducibility and discrimination even in forensic autopsy materials u p to 96 h postmortem. Elevated SP-A expressions were determined in some cas es. This system without post-PCR sample handling would be a very useful too l in pathological diagnosis and DNA analysis. (C) 2000 Elsevier Science Ire land Ltd. All rights reserved.